Contributions
Abstract: EP953
Type: E-Poster Presentation
Session title: Myeloma and other monoclonal gammopathies - Biology & Translational Research
Background
lncRNA is involved in the occurrence, development, metastasis and drug resistance of tumors, and involves a variety of biological functions. In addition，miRNA can regulate proliferation, migration, and even regulate epigenetics to promote the development of MM. However, the mechanism of ncRNA involved in MM is still unclear, and there are many unknown ncRNAs to be explored. Recently, there are more and more researches on the pathogenesis of various tumors with competitive endogenous RNA (ceRNA). ceRNA refers to transcripts that have a common miRNA binding site (MRE) and regulate each other at the post-transcriptional level,which could be used as a potential targeted drug to inhibit tumors. However, how ceRNA plays a role in multiple myeloma (MM) and the mechanism of action of specific ceRNA in the occurrence and development of MM is not fully clear.
Aims
This research aims to discover unknown ncRNAs in MM through high-throughput sequencing, and to study the mechanism and role of ceRNA involved in the pathogenesis of MM for the development of novel molecular markers and potential new targeted drugs.
Methods
In this study, the differentially expressed lncRNA and its target genes were analyzed to predict the lncRNA-miRNA-mRNA interaction pathway though high-throughput sequencing from 8 MM patients and 2 healthy controls. After verifying differential lncRNA and target genes by RT-PCR, lncRNA MSTRG.29039.1 was screened out. We knocked down the expression of MSTRG.29039.1 in MM cell lines by siRNA, detected proliferation by CCK8 as well as apoptosis level by FCM. Besides, we knocked down the miRNA for rescue experiment. The role of OSMR pathway JAK2/STAT3 was verified by Western blot.
Results
262 new lncRNAs were screened by RNA sequencing with statistical differences, 203 were up-regulated and 59 were down-regulated, revealing the characteristics of 63 target genes that were highly expressed in 8 MM patients(Figure1.A、B). According to the function and novelty of the target genes and the expression of lncRNA in MM , we screened out lncRNA MSTRG.29039.1, and verified that it and its target gene OSMR were highly expressed in MM(Figure1. C、D), while the miRNA hsa-miR-12119 was low level in MM(Figure1.E). After knockdown of MSTRG.29039.1 in AMO-1 and U266 MM cell lines, the expression of OSMR was decreased, and the expression of hsa-miR-12119 was up-regulated, which can also promote cell apoptosis and inhibit proliferation. Then,we knocked down hsa-miR-12119 as well as knocked down MSTRG.29039.1 and found that apoptosis was reduced and cell proliferation increased(Figure2.A-F). We further found that MSTRG.29039.1 and OSMR have a common miRNA binding site, and the direct binding relationship was verified by dual-luciferase reporter assay system(Figure3.A-B). Thus, MSTRG.29039.1 can spangy miRNA to counteract the inhibitory effect of miRNA on OSMR. We found that OSMR regulates cell proliferation and apoptosis through the JAK2/STAT3 pathway by Western blot(Figure3.C).
Conclusion
Knockdown of MSTRG.29039.1 could reduce the expression of OSMR to promote apoptosis and inhibit cell proliferation though JAK2/STAT3 pathway, while knockdown of hsa-miR-12119 at the same time could counteract the above phenomenon. This can be a molecular marker and potential therapeutic target for MM.
Keyword(s): Multiple myeloma
Abstract: EP953
Type: E-Poster Presentation
Session title: Myeloma and other monoclonal gammopathies - Biology & Translational Research
Background
lncRNA is involved in the occurrence, development, metastasis and drug resistance of tumors, and involves a variety of biological functions. In addition，miRNA can regulate proliferation, migration, and even regulate epigenetics to promote the development of MM. However, the mechanism of ncRNA involved in MM is still unclear, and there are many unknown ncRNAs to be explored. Recently, there are more and more researches on the pathogenesis of various tumors with competitive endogenous RNA (ceRNA). ceRNA refers to transcripts that have a common miRNA binding site (MRE) and regulate each other at the post-transcriptional level,which could be used as a potential targeted drug to inhibit tumors. However, how ceRNA plays a role in multiple myeloma (MM) and the mechanism of action of specific ceRNA in the occurrence and development of MM is not fully clear.
Aims
This research aims to discover unknown ncRNAs in MM through high-throughput sequencing, and to study the mechanism and role of ceRNA involved in the pathogenesis of MM for the development of novel molecular markers and potential new targeted drugs.
Methods
In this study, the differentially expressed lncRNA and its target genes were analyzed to predict the lncRNA-miRNA-mRNA interaction pathway though high-throughput sequencing from 8 MM patients and 2 healthy controls. After verifying differential lncRNA and target genes by RT-PCR, lncRNA MSTRG.29039.1 was screened out. We knocked down the expression of MSTRG.29039.1 in MM cell lines by siRNA, detected proliferation by CCK8 as well as apoptosis level by FCM. Besides, we knocked down the miRNA for rescue experiment. The role of OSMR pathway JAK2/STAT3 was verified by Western blot.
Results
262 new lncRNAs were screened by RNA sequencing with statistical differences, 203 were up-regulated and 59 were down-regulated, revealing the characteristics of 63 target genes that were highly expressed in 8 MM patients(Figure1.A、B). According to the function and novelty of the target genes and the expression of lncRNA in MM , we screened out lncRNA MSTRG.29039.1, and verified that it and its target gene OSMR were highly expressed in MM(Figure1. C、D), while the miRNA hsa-miR-12119 was low level in MM(Figure1.E). After knockdown of MSTRG.29039.1 in AMO-1 and U266 MM cell lines, the expression of OSMR was decreased, and the expression of hsa-miR-12119 was up-regulated, which can also promote cell apoptosis and inhibit proliferation. Then,we knocked down hsa-miR-12119 as well as knocked down MSTRG.29039.1 and found that apoptosis was reduced and cell proliferation increased(Figure2.A-F). We further found that MSTRG.29039.1 and OSMR have a common miRNA binding site, and the direct binding relationship was verified by dual-luciferase reporter assay system(Figure3.A-B). Thus, MSTRG.29039.1 can spangy miRNA to counteract the inhibitory effect of miRNA on OSMR. We found that OSMR regulates cell proliferation and apoptosis through the JAK2/STAT3 pathway by Western blot(Figure3.C).
Conclusion
Knockdown of MSTRG.29039.1 could reduce the expression of OSMR to promote apoptosis and inhibit cell proliferation though JAK2/STAT3 pathway, while knockdown of hsa-miR-12119 at the same time could counteract the above phenomenon. This can be a molecular marker and potential therapeutic target for MM.
Keyword(s): Multiple myeloma